Shuangyu Tangke Decoction ameliorates diabetic nephropathy by targeting macrophage glycolysis via the Robinin-Coro1a interaction: Insights from multi-omics analysis

Abstract Background Diabetic nephropathy (DN) is a major microvascular complication of diabetes and a leading cause of end-stage renal disease, yet effective therapies remain limited. Shuangyu Tangke Decoction (SKD), a traditional Chinese herbal formula, has shown promising clinical benefits in treating diabetes and DN. Nevertheless, the specific bioactive constituents and underlying molecular mechanisms of SKD remain incompletely understood. Purpose This study aimed to systematically evaluate the renoprotective effects of SKD and elucidate its mechanism of action through integrated multi-omics profiling, computational modeling, and experimental validation. Methods The bioactive serum-absorbed constituents of SKD were characterized using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). The therapeutic efficacy was evaluated in db/db mice through biochemical measurements and renal histopathology. Integrated transcriptomic, proteomic, and metabolomic analyses were utilized to identify the central dysregulated pathway and key molecular targets. Molecular docking, molecular dynamics simulations, microscale thermophoresis (MST), and cellular thermal shift assays (CETSA) were used to validate the interaction between Robinin (a major active constituent) and Coro1a. Functional rescue experiments involving Coro1a overexpression and knockdown in macrophages, along with site-directed mutagenesis, were conducted to identify the critical binding residue. Results SKD reduced blood glucose levels, improved renal function index, and attenuated renal pathological injury in db/db mice. Multi-omics integration identified glycolysis as the central dysregulated pathway, with Coro1a serving as a key molecular node. SKD suppressed macrophage glycolysis, inhibited M1 polarization, and reduced the production of pro-inflammatory cytokines. UPLC-Q-TOF/MS identified 63 prototype compounds and nine serum-absorbed constituents of SKD. Robinin binds to Coro1a at D183, disrupts glycolytic flux, and alleviates inflammation. Conclusion These findings demonstrate that SKD exerts renoprotective effects in DN by targeting macrophage metabolism through inhibition of glycolysis via the Robinin–Coro1a interaction. This study uncovers a mechanism linking herbal medicine to immune-metabolic crosstalk and identifies Coro1a as a potential therapeutic target for metabolic kidney diseases. Our work provides a mechanistic foundation for the clinical application of SKD and advances the evidence-based development of plant-derived therapeutics for DN.